Date of Award

8-2025

Document Type

Thesis

Degree Name

Master of Science (MS)

College/School

College of Science and Mathematics

Department/Program

Chemistry and Biochemistry

Thesis Sponsor/Dissertation Chair/Project Chair

David Rotella

Committee Member

Jim Dyer

Committee Member

Nina Goodey

Abstract

Malaria and Toxoplasmosis are life threatening diseases to humans. The search for a cure and control of these diseases is still one of the major challenges of scientists worldwide. In this research we focused on two Malaria species; Plasmodium bergei² (Pb) and Plasmodium falciparum¹ (Pf) and also on Toxoplasma gondii. Part of the life cycle of these organisms is regulated by the enzyme cyclic GMP-dependent protein kinase G (PKG)⁴. This is our drug target in this research. We believe that inhibiting this enzyme will be a great step in fighting these diseases. In this research, we devised a comparative study on cloning and producing this enzyme in two separate systems; eukaryotic system (Sf9 insect cells) and Escherichia coli (prokaryotic system for Pf and Pb PKG ). To do this, the process involved cloning of the genes and transformation of cells, protein expression, His-tag purification, characterization (cGMP and ATP dependency), kinetic studies, and inhibition profiles by a known PKG inhibitor (trisubstituted pyrrole TSP). After doing these, we came to a conclusion that the Sf9 insect cell system will be a great way forward to produce this enzyme in significant amounts.

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